C O M M U N I C A T I O N S
Table 1. Activities of Colchicine Neoglyosides
a The saccharide portion of the library member is represented. b Cytoxicity (Xb5M-1) as determined by cell titer-glo and calcein AM assays (see Supporting
Information for assay parameters). c The results of tubulin polymerization assays where “D” designates destabilizer and “S” designates stabilizer (see Supporting
Information for assay parameters). d Synergism or antagonism in drug combination studies with the parent 1 (a representative destabilizer) or paclitaxel (a
representative stabilizer) analyzed via the Chou-Talalay method (see ref 11). The results of synergy assays-legend (combination index): (++++) strong
synergism, CI 0.1-0.3; (+++) synergism, CI 0.3-0.7; (++) moderate synergism, CI 0.7-0.85; (+) slight synergism, CI 0.85-0.9; (-) slight antagonism,
CI 1.1-1.2; (--) moderate antagonism, CI 1.2-1.45; (---) antagonism, CI 1.45-3.3; (----) strong antagonism, CI 3.3-10. e Library member contains both
pyranose and furanose forms (see ref 12).
from 22 to 35 nM), as did neoglycosides Col19 and Col21, albeit
both were roughly 1 order of magnitude less potent than 1. It should
References
(1) (a) Thorson, J. S.; Vogt, T. In Carbohydrate-Based Drug DiscoVery;
be noted that, while the best neoglycosides (Col19 and Col21)
Wong, C. H., Ed.; Wiley-VCH: Weinheim, Germany, 2003; p 685. (b)
Kren, V.; Martinkova, L. Curr. Med. Chem. 2001, 8, 1303. (c) Thorson,
J. S.; Hosted, T. J., Jr.; Jiang, J.; Biggins, J. B.; Ahlert, J. Curr. Org.
Chem. 2001, 5, 139. (d) Weymouth-Wilson, A. C. Nat. Prod. Rep. 1997,
14, 99.
displayed roughly a 10-fold reduction in potency, the IC50 values
of these colchicine neoglycosides still fall within the range of the
clinically relevant cytotoxins doxorubicin and paclitaxel.
(2) (a) Griffith, B. R.; Langenhan, J. M.; Thorson, J. S. Curr. Opin. Biotechnol.
2005, 16, 622. (b) Langenhan, J. M.; Griffith, B. R.; Thorson, J. S. J.
Nat. Prod. 2005, 68, 1696. (c) Yang, J.; Hoffmeister, D.; Liu, L.; Fu, X.;
Thorson, J. S. Bioorg. Med. Chem. 2004, 12, 1577.
(3) (a) Zhang, C.; Griffith, B. R.; Fu, Q.; Albermann, C.; Fu, X.; Lee, I.-K.;
Li, L.; Thorson, J. S. Science 2006, 313, 1291. (b) Borisova, S. A.; Zhang,
C.; Takahashi, H.; Zhang, H.; Wong, A. W.; Thorson, J. S.; Liu, H.-w.
Angew. Chem., Intl. Ed. 2006, 45, 2748. (c) Fu, X.; Albermann, C.; Zhang,
C.; Thorson, J. S. Org. Lett. 2005, 7, 1513. (d) Albermann, C.; Soriano,
A.; Jiang, J.; Vollmer, H.; Biggins, J. B.; Barton, W. A.; Lesniak, J.;
Nikolov, D. B.; Thorson, Jon S. Org. Lett. 2003, 5, 933. (e) Fu, X.;
Albermann, C.; Jiang, J.; Liao, J.; Zhang, C.; Thorson, J. S. Nat.
Biotechnol. 2003, 21, 1467.
(4) Langenhan, J. M.; Peters, N. R.; Guzei, I. A.; Hoffmann, F. M.; Thorson,
J. S. Proc. Natl. Acad. Sci. U.S.A. 2005, 102, 12305.
(5) (a) Graening, T.; Schmalz, H. G. Angew. Chem., Int. Ed. 2004, 43, 3230
and references therein. (b) Wipf, P.; Reeves, J. T.; Day, B. W. Curr.
Pharm. Des. 2004, 10, 1417. (c) Timasheff, S.; Andreu, J.; Gorbunoff,
M.; Medranot, F.; Prakash, V. Cell. Pharmacol. 1993, 1, S27. (d) Muzaffar,
A.; Brossi, A. Pharmacol. Ther. 1991, 49, 105.
(6) (a) Buskila, D.; Zaks, N.; Neumann, L.; Livneh, A.; Greenberg, S.; Pras,
M.; Langevitz, P. Clin. Exp. Rheumatol. 1997, 15, 355. (b) Levy, M.;
Spino, M.; Read, S. Pharmacotherapy 1991, 11, 196.
(7) (a) Iacobuzio-Donahue, C. A.; Lee, E. L.; Abraham, S. C.; Yardley, J.
H.; Wu, T. T. Am. J. Surg. Pathol. 2001, 25, 1067. (b) Kubler, P. A.
Med. J. Aust. 2000, 172, 498. (c) Simons, R. J.; Kingma, D. W. Am. J.
Med. 1989, 86, 356. (d) Luciani, I. J. Emerg. Nurs. 1989, 15, 80. (e)
Stanley, M. W.; Taurog, J. D.; Snover, D. C. Clin. Exp. Rheumatol. 1984,
2, 167.
(8) Bombardelli, E.; Fontana, G. PCT Int. Appl. WO 2004111068 A1, 2004.
(9) Bagnato J. D.; Eilers A. L.; Horton R. A.; Grissom C. B. J. Org. Chem.
2004, 69, 8987.
(10) Colchicine-D-neoglucoside (Col0): see Supporting Information for ex-
perimental details and characterization data.
To assess how these structural modifications affect the ability
of library members to modulate tubulin polymerization, a funda-
mental activity of 1, the same 15 “hits” were submitted to a
secondary in vitro tubulin polymerization assay.13 As expected, eight
compounds (including Col6 and Col45, Table 1) exhibited effects
on microtubules consistent with the destabilizing effects of 1.
However, three compounds (including Col56 and Col65, Table 1)
had no apparent effect on tubulin polymerization (similar to the
standard doxorubicin), and surprisingly, two compounds (Col19
and Col21, Table 1) exhibited effects on microtubules consistent
with the stabilizing effects of paclitaxel. Drug combination assays
were subsequently performed to determine if the mechanism of
action of tubulin binding by 1 had been affected for compounds
Col19 and Col21.11 Consistent with the in vitro tubulin polymer-
ization results, Col19 and Col21 showed synergism with the parent
1 and antagonism with paclitaxel, suggesting the Col19/21-tubulin
interaction to mirror that of paclitaxel-tubulin. While many
synthetic and natural product small molecules are known to stabilize
or destabilize tubulin polymerization,14 to the best of our knowledge,
this stands as the first example of interconverting these two distinct
mechanisms via simple synthetic derivatization. Cumulatively, this
study highlights a simple extension of neoglycorandomization
toward amine-bearing scaffolds and illustrates a potential benefit
to glycosylating nonglycosylated natural products.
Acknowledgment. We thank the University of Wisconsins
Madison School of Pharmacy Analytical Instrumentation Facility
for analytical support and Dr. Byron R. Griffith, Prof. C. Richard
Hutchinson, and Dr. Thomas Stringfellow for helpful discussion.
This research was supported in part by National Institutes of Health
Grants U19 CA113297, AI552218, CA84374, and GM70637. J.S.T.
is a HI Romnes Fellow.
Supporting Information Available: Experimental procedures,
compound/library characterization data, complete cytotoxicity, and
tubulin polymerization assay data. This material is available free of
(11) (a) Chou T.-C.; Hayball, M. P. Dose effect analysis; software and manual;
Biosoft: Cambridge, U.K., 1996. (b) Chou, T.-C.; Talalay, P. AdV. Enzyme
Regul. 1984, 22, 27.
(12) (a) The isomeric distribution of pyranose and furanose neoglyosides is
dependent upon the identity of the sugar (see Peri, F.; Dumy, P.; Mutter,
M.; Tetrahedron 1998, 54, 12269 and ref 4) and the specific mutarotation
of 2-deoxy ribose has been reported (see Lemieux, R. U.; Anderson, L.;
Conner, A. H. Carbohydr. Res. 1971, 20, 59). Col21: see Supporting
Information for experimental details and characterization data.
(13) Bonne, D.; Heusele, C.; Simon, C.; Pantaloni, D. J. Biol. Chem. 1985,
260, 2819.
(14) (a) Jordan, M. A.; Wilson, L. Nat. ReV. Cancer 2004, 4, 253. (b) Islam,
M. N.; Iskander, M. N. Mini-ReV. Med. Chem. 2004, 4, 1077. (c) Von A.
E. Exp. Opin. Ther. Patent. 1999, 9, 1069. (d) Jordan, A.; Hadfield, J.
A.; Lawrence, N. J.; McGown, A. T. Med. Res. ReV. 1998, 18, 259.
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